Anti-TIGIT and anti-CD96 antibodies

Abstract

The instant disclosure provides multispecific molecules that specifically bind to CD96 (e.g., human CD96) and/or TIGIT (e.g., human TIGIT) and isolated antibodies that specifically bind to TIGIT (e.g., human TIGIT). Also provided are pharmaceutical compositions comprising these multispecific molecules and antibodies, nucleic acids encoding these multispecific molecules and antibodies, expression vectors and host cells for making these multispecific molecules and antibodies, and methods of treating a subject using these multispecific molecules and antibodies.

What is claimed: 
     
       1. A multispecific molecule comprising:
 (a) a first antigen-binding region that specifically binds to human CD96, the first antigen-binding region comprising a first heavy chain variable region (VH) comprising CDRs CDRH1, CDRH2, and CDRH3, and a first light chain variable region (VL) comprising CDRs CDRL1, CDRL2, and CDRL3; wherein the first VH comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence of SEQ ID NO: 38; and the first VL comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence of SEQ ID NO: 39; and 
 (b) a second antigen-binding region that specifically binds to human TIGIT, the second antigen-binding region comprising a second VH comprising CDRs CDRH1, CDRH2, and CDRH3, and a second VL comprising CDRs CDRL I, CDRL2, and CDRL3; wherein the second VH comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence of SEQ ID NO: 40; and/or the second VL comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence of SEQ ID NO: 41. 
 
     
     
       2. A multispecific molecule comprising:
 (a) a first antigen-binding region that specifically binds to human CD96, comprising a first VH comprising CDRs CDRH1, CDRH2, and CDRH3, and a first VL comprising CDRs CDRL1, CDRL2, and CDRL3, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the first antigen-binding region comprise the amino acid sequences of SEQ ID NOs: 22, 23, 24, 25, 26, and 27, respectively; and 
 (b) a second antigen-binding region that specifically binds to human TIGIT, comprising a second VH comprising CDRs CDRH1, CDRH2, and CDRH3, and a second VL comprising CDRs CDRL1, CDRL2, and CDRL3, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the second antigen-binding region comprise the amino acid sequences of SEQ ID NOs: 28, 29, 30, 31, 32, and 33, respectively. 
 
     
     
       3. The multispecific molecule of  claim 2 , wherein:
 (a) the first VH comprises an amino acid sequence at least 95% identical to SEQ ID NO: 38; 
 (b) the first VH comprises the amino acid sequence of SEQ ID NO: 38; 
 (c) the first VL comprises an amino acid sequence at least 95% identical to SEQ ID NO: 39; 
 (d) the first VL comprises the amino acid sequence of SEQ ID NO: 39; 
 (e) the first VH and first VL comprise the amino acid sequences of SEQ ID NOs: 38 and 39, respectively; 
 (f) the second VH comprises an amino acid sequence at least 95% identical to SEQ ID NO: 40; 
 (g) the second VH comprises the amino acid sequence of SEQ ID NO: 40; 
 (h) the second VL comprises an amino acid sequence at least 95% identical to SEQ ID NO: 41; 
 (i) the second VL comprises the amino acid sequence of SEQ ID NO: 41; and/or 
 (j) the second VH and second VL comprise the amino acid sequences of SEQ ID NOs: 40 and 41, respectively. 
 
     
     
       4. The multispecific molecule of  claim 2 , wherein the first antigen-binding region and/or the second antigen-binding region comprises a heavy chain constant region selected from the group consisting of human IgG 1 , IgG 2 , IgG 3 , IgG 4 , IgA 1 , and IgA 2 . 
     
     
       5. The multispecific molecule of  claim 2 , wherein:
 (a) the first antigen-binding region comprises a first human IgG heavy chain constant region comprising aspartate, serine, alanine, and valine at amino acid positions 239, 366, 368, and 407, respectively, and the second antigen-binding region comprises a second human IgG heavy chain constant region comprising aspartate, leucine, glutamate, and tryptophan at amino acid positions 239, 330, 332, and 366, respectively; and/or 
 (b) the first antigen-binding region comprises a first human IgG heavy chain constant region comprising aspartate, leucine, glutamate, and tryptophan at amino acid positions 239, 330, 332, and 366, respectively, and the second antigen-binding region comprises a second human IgG heavy chain constant region comprising aspartate, serine, alanine, and valine at amino acid positions 239, 366, 368, and 407, respectively, 
 wherein the amino acid positions are numbered according to the EU numbering system. 
 
     
     
       6. The multispecific molecule of  claim 5 , wherein:
 (a) the first antigen-binding region comprises a first heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 50, and the second antigen-binding region comprises a second heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 56; and/or 
 (b) the first antigen-binding region comprises a first heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 56, and the second antigen-binding region comprises a second heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 50. 
 
     
     
       7. The multispecific molecule of  claim 2 , wherein the first antigen-binding region comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 5, and/or the second antigen-binding region comprises a second heavy chain comprising the amino acid sequence of SEQ ID NO: 7. 
     
     
       8. The multispecific molecule of  claim 2 , wherein the first antigen-binding region and/or the second antigen-binding region comprises a human kappa light chain constant region or a human lambda light chain constant region. 
     
     
       9. The multispecific molecule of  claim 2 , wherein the first antigen-binding region comprises a first light chain comprising the amino acid sequence of SEQ ID NO: 6, and/or the second antigen-binding region comprises a second light chain comprising the amino acid sequence of SEQ ID NO: 9. 
     
     
       10. The multispecific molecule of  claim 2 , wherein:
 (a) the first VH comprises the amino acid sequence of SEQ ID NO: 38, and the first VL comprises the amino acid sequence of SEQ ID NO: 39; and 
 (b) the second VH comprises the amino acid sequence of SEQ ID NO: 40, and the second VL comprises the amino acid sequence of SEQ ID NO: 41. 
 
     
     
       11. The multispecific molecule of  claim 10 , wherein:
 (a) the first antigen-binding region comprises a first heavy chain comprising an amino acid sequence at least 95% identical to SEQ ID NO: 5; 
 (b) the second antigen-binding region comprises a second heavy chain comprising an amino acid sequence at least 95% identical to SEQ ID NO: 7; 
 (c) the first antigen-binding region comprises a first light chain comprising an amino acid sequence at least 95% identical to SEQ ID NO: 6; and/or 
 (d) the second antigen-binding region comprises a second light chain comprising an amino acid sequence at least 95% identical to SEQ ID NO: 9. 
 
     
     
       12. The multispecific molecule of  claim 2  wherein:
 (a) the first antigen-binding region comprises a first VH comprising an amino acid sequence at least 95% identical to SEQ ID NO:38; 
 (b) the first antigen-binding region comprises a first VL comprising an amino acid sequence at least 95% identical to SEQ ID NO:39; 
 (c) the second antigen-binding region comprises a second VH comprising an amino acid sequence at least 95% identical to SEQ ID NO:40; and/or (d) the second antigen-binding region comprises a first VL comprising an amino acid sequence at least 95% identical to SEQ ID NO:41. 
 
     
     
       13. The multispecific molecule of  claim 10 , wherein the first antigen-binding region and/or the second antigen-binding region comprises a heavy chain constant region selected from the group consisting of human IgG 1 , IgG 2 , IgG 3 , IgG 4 , IgA 1 , and IgA 2 . 
     
     
       14. The multispecific molecule of  claim 13 , wherein:
 (a) the first antigen-binding region comprises a first human IgG heavy chain constant region comprising aspartate, serine, alanine, and valine at amino acid positions 239, 366, 368, and 407, respectively, and the second antigen-binding region comprises a second human IgG heavy chain constant region comprising aspartate, leucine, glutamate, and tryptophan at amino acid positions 239, 330, 332, and 366, respectively; and/or 
 (b) the first antigen-binding region comprises a first human IgG heavy chain constant region constant region comprising aspartate, leucine, glutamate, and tryptophan at amino acid positions 239, 330, 332, and 366, respectively, and the second antigen-binding region comprises a second human IgG heavy chain constant region comprising aspartate, serine, alanine, and valine at amino acid positions 239, 366, 368, and 407, respectively, 
 wherein the amino acid positions are numbered according to the EU numbering system. 
 
     
     
       15. The multispecific molecule of  claim 14 , wherein:
 (a) the first antigen-binding region comprises a first heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 50, and the second antigen-binding region comprises a second heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 56; and/or 
 (b) the first antigen-binding region comprises a first heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 56, and the second antigen-binding region comprises a second heavy chain constant region comprising the amino acid sequence of SEQ ID NO: 50. 
 
     
     
       16. The multispecific molecule of  claim 10 , wherein the first antigen-binding region comprises a first heavy chain comprising the amino acid sequence of SEQ ID NO: 5, and/or the second antigen-binding region comprises a second heavy chain comprising the amino acid sequence of SEQ ID NO: 7. 
     
     
       17. The multispecific molecule of  claim 10 , wherein the first antigen-binding region and/or the second antigen-binding region comprises a human kappa light chain constant region or a human lambda light chain constant region. 
     
     
       18. The multispecific molecule of  claim 10 , wherein the first antigen-binding region comprises a first light chain comprising the amino acid sequence of SEQ ID NO: 6, and/or the second antigen-binding region comprises a second light chain comprising the amino acid sequence of SEQ ID NO: 9. 
     
     
       19. A multispecific molecule comprising:
 (a) a first antigen-binding region that specifically binds to human CD96, the first antigen-binding region comprising a first heavy chain comprising the amino acid sequence of SEQ ID NO: 5, and a first light chain comprising the amino acid sequence of SEQ ID NO: 6; and 
 (b) a second antigen-binding region that specifically binds to human TIGIT, the second antigen-binding region comprising a second heavy chain comprising the amino acid sequence of SEQ ID NO: 7, and a second light chain comprising the amino acid sequence of SEQ ID NO: 9. 
 
     
     
       20. An antibody that specifically binds to human TIGIT, the antibody comprising: a VH comprising the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence of SEQ ID NO: 40; and a VL comprising the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence of SEQ ID NO: 41. 
     
     
       21. The antibody of  claim 20 , comprising the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 amino acid sequences of SEQ ID NOs: 28, 29, 30, 31, 32, and 33, respectively. 
     
     
       22. The antibody of  claim 20 , comprising the VH and VL amino acid sequences of SEQ ID NOs: 40 and 41, respectively. 
     
     
       23. The antibody of  claim 20 , wherein the antibody comprises a heavy chain constant region selected from the group consisting of human IgG 1 , IgG 2 , IgG 3 , IgG 4 , IgA 1 , and IgA 2 . 
     
     
       24. The antibody of  claim 23 , wherein the heavy chain constant region comprises the amino acid sequence of any one of SEQ ID NO: 49-60. 
     
     
       25. The antibody of  claim 20 , the antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 7, and a light chain comprising the amino acid sequence of SEQ ID NO: 9. 
     
     
       26. A multispecific molecule comprising:
 (a) a first antigen-binding region that specifically binds to human CD96, the first antigen-binding region comprising a first VH comprising CDRs CDRH1, CDRH2, and CDRH3, and a first VL comprising CDRs CDRL1, CDRL2, and CDRL3, wherein the first VH comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence of SEQ ID NO: 38; and the first VL comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence of SEQ ID NO: 39; and 
 (b) a second antigen-binding region that specifically binds to human TIGIT. 
 
     
     
       27. The multispecific molecule of  claim 26 , wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the first antigen-binding region comprise the amino acid sequences of SEQ ID NOs: 22, 23, 24, 25, 26, and 27, respectively. 
     
     
       28. The multispecific molecule of  claim 27 , wherein the first VH and first VL comprise the amino acid sequences of SEQ ID NOs: 38 and 39, respectively. 
     
     
       29. The multispecific molecule of  claim 27 , wherein the first antigen-binding region and/or the second antigen-binding region comprises a heavy chain constant region selected from the group consisting of human IgG 1 , IgG 2 , IgG 3 , IgG 4 , IgA 1 , and IgA 2 . 
     
     
       30. The multispecific molecule of  claim 29 , wherein the heavy chain constant region comprises the amino acid sequence of any one of SEQ ID NO: 49-60. 
     
     
       31. The multispecific molecule of  claim 27 , the first antigen-binding region comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 5, and a light chain comprising the amino acid sequence of SEQ ID NO: 6. 
     
     
       32. A multispecific molecule comprising:
 (a) a first antigen-binding region that specifically binds to human CD96; and 
 (b) a second antigen-binding region that specifically binds to human TIGIT, the second antigen-binding region comprising a VH comprising CDRs CDRH1, CDRH2, and CDRH3 and a VL comprising CDRs CDRL1, CDRL2, and CDRL3, wherein the VH comprises the CDRH1, CDRH2, and CDRH3 amino acid sequences of the VH amino acid sequence of SEQ ID NO: 40; 
 and the VL comprises the CDRL1, CDRL2, and CDRL3 amino acid sequences of the VL amino acid sequence of SEQ ID NO: 41. 
 
     
     
       33. The multispecific molecule of  claim 32 , wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 of the second antigen-binding region comprise the amino acid sequences of SEQ ID NOs: 28, 29, 30, 31, 32, and 33, respectively. 
     
     
       34. The multispecific molecule of  claim 32 , wherein the VH and VL of the second antigen-binding region comprise the amino acid sequences of SEQ ID NOs: 40 and 41, respectively. 
     
     
       35. The multispecific molecule of  claim 32 , wherein the first antigen-binding region and/or the second antigen-binding region comprises a heavy chain constant region selected from the group consisting of human IgG 1 , IgG 2 , IgG 3 , IgG 4 , IgA 1 , and IgA 2 . 
     
     
       36. The multispecific molecule of  claim 35 , wherein the heavy chain constant region comprises the amino acid sequence of any one of SEQ ID NO: 49-60. 
     
     
       37. The multispecific molecule of  claim 32 , wherein the second antigen-binding region comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 7, and a light chain comprising the amino acid sequence of SEQ ID NO: 9.